Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2
ABSTRACTPseudoviruses are useful virological tools because of their safety and versatility, especially for emerging and re-emerging viruses. Due to its high pathogenicity and infectivity and the lack of effective vaccines and therapeutics, live SARS-CoV-2 has to be handled under biosafety level 3 co...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-01-01
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| Series: | Emerging Microbes and Infections |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2020.1743767 |
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| author | Jianhui Nie Qianqian Li Jiajing Wu Chenyan Zhao Huan Hao Huan Liu Li Zhang Lingling Nie Haiyang Qin Meng Wang Qiong Lu Xiaoyu Li Qiyu Sun Junkai Liu Changfa Fan Weijin Huang Miao Xu Youchun Wang |
| author_facet | Jianhui Nie Qianqian Li Jiajing Wu Chenyan Zhao Huan Hao Huan Liu Li Zhang Lingling Nie Haiyang Qin Meng Wang Qiong Lu Xiaoyu Li Qiyu Sun Junkai Liu Changfa Fan Weijin Huang Miao Xu Youchun Wang |
| author_sort | Jianhui Nie |
| collection | DOAJ |
| description | ABSTRACTPseudoviruses are useful virological tools because of their safety and versatility, especially for emerging and re-emerging viruses. Due to its high pathogenicity and infectivity and the lack of effective vaccines and therapeutics, live SARS-CoV-2 has to be handled under biosafety level 3 conditions, which has hindered the development of vaccines and therapeutics. Based on a VSV pseudovirus production system, a pseudovirus-based neutralization assay has been developed for evaluating neutralizing antibodies against SARS-CoV-2 in biosafety level 2 facilities. The key parameters for this assay were optimized, including cell types, cell numbers, virus inoculum. When tested against the SARS-CoV-2 pseudovirus, SARS-CoV-2 convalescent patient sera showed high neutralizing potency, which underscore its potential as therapeutics. The limit of detection for this assay was determined as 22.1 and 43.2 for human and mouse serum samples respectively using a panel of 120 negative samples. The cutoff values were set as 30 and 50 for human and mouse serum samples, respectively. This assay showed relatively low coefficient of variations with 15.9% and 16.2% for the intra- and inter-assay analyses respectively. Taken together, we established a robust pseudovirus-based neutralization assay for SARS-CoV-2 and are glad to share pseudoviruses and related protocols with the developers of vaccines or therapeutics to fight against this lethal virus. |
| first_indexed | 2024-03-07T17:24:03Z |
| format | Article |
| id | doaj.art-7449af54efaa49489c65e8cffb9a8750 |
| institution | Directory Open Access Journal |
| issn | 2222-1751 |
| language | English |
| last_indexed | 2024-04-25T00:51:44Z |
| publishDate | 2020-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj.art-7449af54efaa49489c65e8cffb9a87502024-03-11T16:04:24ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512020-01-019168068610.1080/22221751.2020.1743767Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2Jianhui Nie0Qianqian Li1Jiajing Wu2Chenyan Zhao3Huan Hao4Huan Liu5Li Zhang6Lingling Nie7Haiyang Qin8Meng Wang9Qiong Lu10Xiaoyu Li11Qiyu Sun12Junkai Liu13Changfa Fan14Weijin Huang15Miao Xu16Youchun Wang17Division of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaDivision of Animal Model Research, Institute for Laboratory Animal Resources, National Institutes for Food and Drug Control, Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaInstitute for Biological Product Control, National Institutes for Food and Drug Control, Beijing, People’s Republic of ChinaDivision of HIV/AIDS and Sex-transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing, People’s Republic of ChinaABSTRACTPseudoviruses are useful virological tools because of their safety and versatility, especially for emerging and re-emerging viruses. Due to its high pathogenicity and infectivity and the lack of effective vaccines and therapeutics, live SARS-CoV-2 has to be handled under biosafety level 3 conditions, which has hindered the development of vaccines and therapeutics. Based on a VSV pseudovirus production system, a pseudovirus-based neutralization assay has been developed for evaluating neutralizing antibodies against SARS-CoV-2 in biosafety level 2 facilities. The key parameters for this assay were optimized, including cell types, cell numbers, virus inoculum. When tested against the SARS-CoV-2 pseudovirus, SARS-CoV-2 convalescent patient sera showed high neutralizing potency, which underscore its potential as therapeutics. The limit of detection for this assay was determined as 22.1 and 43.2 for human and mouse serum samples respectively using a panel of 120 negative samples. The cutoff values were set as 30 and 50 for human and mouse serum samples, respectively. This assay showed relatively low coefficient of variations with 15.9% and 16.2% for the intra- and inter-assay analyses respectively. Taken together, we established a robust pseudovirus-based neutralization assay for SARS-CoV-2 and are glad to share pseudoviruses and related protocols with the developers of vaccines or therapeutics to fight against this lethal virus.https://www.tandfonline.com/doi/10.1080/22221751.2020.1743767SARS-CoV-2COVID-19neutralizing antibodypseudovirusneutralization assay |
| spellingShingle | Jianhui Nie Qianqian Li Jiajing Wu Chenyan Zhao Huan Hao Huan Liu Li Zhang Lingling Nie Haiyang Qin Meng Wang Qiong Lu Xiaoyu Li Qiyu Sun Junkai Liu Changfa Fan Weijin Huang Miao Xu Youchun Wang Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 Emerging Microbes and Infections SARS-CoV-2 COVID-19 neutralizing antibody pseudovirus neutralization assay |
| title | Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 |
| title_full | Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 |
| title_fullStr | Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 |
| title_full_unstemmed | Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 |
| title_short | Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2 |
| title_sort | establishment and validation of a pseudovirus neutralization assay for sars cov 2 |
| topic | SARS-CoV-2 COVID-19 neutralizing antibody pseudovirus neutralization assay |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2020.1743767 |
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